Stable natural color process, products and use thereof

ABSTRACT

A method of preparing colored products from edible materials comprises processing Genipa americana fruit juice, which contains genipin, genipin derivatives, or pre-genipin compounds, with other edible juices or extracts which contain nitrogenous compounds such as amino acids, polypeptides, or proteins. The generated natural colored products have excellent stability and can be used in a broad range of applications including beverages, foodstuffs, drugs, dietary supplements, cosmetics, personal care stuffs, and animal feeds.

This application is related to and claims priority from U.S. ProvisionalApplication Ser. No. 61/040,208, Wu et al, filed Mar. 28, 2008,incorporated herein by reference.

TECHNICAL FIELD

This disclosure relates to natural stable color products which can beused in a broad range of applications including foodstuffs, drugs,nutritional supplements, personal care stuffs, cosmetics, and animalfeeds, and a process for preparing these colored food and edibleproducts. Specifically, the disclosure relates to the stable colorproducts that are generated by processing Genipa americana fruit juicewhich contains genipin, genipin derivatives, or pre-genipin compounds,together with other edible juices or extracts which contain amino acids,polypeptides, proteins, and compounds with one or more primary aminegroups.

BACKGROUND

Today, a decreasing number of people are willing to eat or drinkfoodstuffs and use cosmetic products which are colored synthetically,resulting in a steady growth of the market for natural colorants. Thus,a natural color, especially a natural blue colorant that is stable atlow pH and to temperatures used in processing, would be of significantworldwide commercial interest. The only natural blue colors commerciallyfeasible today are those derived from gardenia fruits (Gardeniajasminoides Ellis). However gardenia blue is not currently available inUS and Europe markets, it is only available in some Asian markets as asafe color product for food/drug applications.

Gardenia fruit contains a large amount of iridoids such as geniposide,gardenoside, genipin-1-b-gentiobioside, geniposidic acid and genipin(Endo, T. and Taguchi, H. Chem. Pharm. Bull. 1973). Among them genipinis a key compound contributing to the gardenia blue whenever it reactswith amino acids (U.S. Pat. No. 4,878,921). Currently gardenia blue ismade as a chemical reaction product using geniposide extracted fromGardenia juice, purified genipin, or genipin derivatives, with isolatedamino acids. In contrast, the process of the present invention utilizeswhole fruit, puree or juice of genipin-containing plants to provide anatural color juice or concentrate.

Genipin and other iridoid compounds, such as genipic acid, genipingentiobioside, geniposide and geniposidic acid are found also in thefruits of Genipa americana, also known as Genipap, or Huito, a wildplant of tropical Latin American. The mature fruits of Genipa americanahave been commonly used by local people to make cooling drinks, jellies,sherbet, ice cream, sweet preserves, syrup, a soft drink genipapada,wine, a potent liqueur, and tanning extract. Green or unripe fruits areused as a color source to paint faces and bodies for adornment, to repelinsects, and to dye clothing, pottery, hammocks, utensils and basketmaterials a bluish-purple. The fruit and juices also have medicinalproperties and syrups are used as cold and cough medicine. Flowers andbark of Genipa americana also have medicinal properties.

Genipa americana is also a natural source of iron, riboflavin andanti-bacterial substances, apart from the carbohydrates, sugar,proteins, ash and malic acid in its fruits. The principal biochemicalcompounds of Genipa americana include: calcium, phosphorous, vitamin C,and caffeine, caterine, genipic acid, genipin, genipin gentiobioside,genipinic acid, geniposide, geniposidic acid, gardenoside, genamesidesA-D, gardendiol, deacetyl asperulosidic acid methyl ester, shanzhiside,glycerides, hydantoin, mannitol, methyl esters, tannic and tartaricacid, and tannins.

The present disclosure relates to the preparation of stable colorants bymixing and co-processing fruit juice, particularly from Genipaamericana, with other edible juices or extracts from fruit, vegetable,plant materials, grain, legume, nuts, seeds, animal materials includingmilk and egg, microbial, and algal materials, which contain amino acids,or polypeptides, or proteins. The product colors are not those expectedfrom simple pigment blending, and the color products have greatstability to acidity and heating. The co-processing of Genipa americanafruit with other selected fruit(s) (defined broadly as above to includealso grains and animal materials) to obtain the composite juice issimple and effective. The products can be used in a broad range ofapplications, such as foodstuffs, drugs, nutritional supplements,personal care stuffs, cosmetics, and animal feed.

SUMMARY OF THE INVENTION

The present disclosure provides a method of producing natural stablecolor products wherein Genipa americana fruits are co-processed withother edible juices or extracts from fruits, vegetables, plantmaterials, grains, legumes, nuts, seeds, animal materials including milkand eggs, microbial, and algal materials that contain amino acids,polypeptides, and/or proteins.

The present disclosure further provides natural color products that havedesirable organoleptic properties and excellent compatibility with foodsand food components, are intrinsically acceptable to consumers due tothe nature of their raw materials and have certain nutritional values,and which also have very good stability over a broad pH range and goodresistance in use to heat and light. Therefore, the products aresuitable for use in various applications, such as foodstuffs, drugs,nutritional supplements, personal care stuffs, cosmetic, and animalfeed.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a flow chart representation of the process described inExample 3.

DETAILED DESCRIPTION OF THE INVENTION

The present disclosure provides a method of producing natural stablecolor products by using Genipa americana fruit juice with the variousadded edible materials. Natural stable blue color is believed to beobtained when genipin and possibly genipin gentiobioside, whichnaturally exist in Genipa americana fruit, react with amino acids,polypeptides, or proteins, and other compounds with primary aminegroups, in the various edible materials. All known and practiced juiceextraction techniques and operations, and processing technologiesassociated with non juice components claimed, are considered suitable toprepare the color juices of this invention and are incorporated byreference herein.

Starting Materials

The starting materials used in the process of the present invention aremature fruits of Genipa americana L.; also known by numerous informalnames: genipap, huito, jaguar, bilito, cafecillo denta, caruto, carutorebalsero, confiture de singe, danipa, genipa, genipa, genipayer bitu,guaitil, guaricha, guayatil colorado, huitol, huitoc, huitu, irayol,jagua blanca, jagua amarilla, jagua colorado, jeipapeiro, juniper,maluco, mandipa, marmelade-box, nandipa, ñandipa genipapo, tapaculo,tapoeripa, taproepa totumillo, yagua, yanupa-i, yenipa-i, yenipapa bi,genipapo, huitoc, vito, chipara, guanapay, or other varieties such asjenipaporana or jenipapobravo. The fruit is optimal for harvest whenmature in size, firm, and green to greenish brown in color; overly ripefruits fall to the ground and decay.

Materials may be whole fruit, fruit juice, fruit puree, fruit juiceconcentrate, dried powder from fruits or juice, and the water-insolublepart of fruits from Genipa americana L or its close relatives that alsocontain genipin, genipin derivatives, or pre-genipin compounds. Thesecompounds are a subgroup of the iridoid class of phytochemicals that arewidely distributed, including in the Rubiaceae family. Some attentionhas been paid to an understanding of iridoid biosynthesis as an elementof chemosystematics (Sampaio-Santos and Kaplan, J. Braz. Chem. Soc. 12(2001)), and these data help to delineate the group ofgenipin-containing plants that produce fruit and juice suitable for useaccording to this invention. Generally, such fruits would includemembers of the Rubiaceae family, which contains sufficient genipin orgenipin derivatives, including genipin gentiobioside, geniposide,geniposidic acid and gardenoside, to react with the other compoundsdefined herein to produce a colored product. It has been reported on thebasis of DNA data (Andreason and Bremer, Am. J. Botany 87 1731-1748(2000)) that Genipa americana comprises a clade together with Gardeniaand Kailarsenia. Without restricting the scope of fruits that may beused in the present invention, mention of Genipa americana should betaken to include edible fruits or juices or extracts or edible parts ofrelated plants such as Gardenia and Kailarsenia and othergenipin-containing plants, including Gardenia jasminoides Ellis and itsvarieties.

Materials may be aqueous or solvent extracts from the sources described.Solvents can be those of common use in the art, including water, acetic,citric, or phosphate buffer solutions, methanol, ethanol, isopropanol,or mixtures thereof in different ratios.

The broad range of suitable edible materials comprises fruits,vegetables, grains, legumes, nuts, seeds, plant materials, animalmaterials including milk and eggs, microbial and algal materials, andby-products from such sources, which contain amino acids, polypeptides,and proteins. These materials can be co-processed with the fruit orfruit juice of Genipa americana to produce natural stable colorants,especially blue color.

The co-process fruits, fruit juice, puree, juice concentrate, driedpowder or extracts may be obtained from fruits included within thegroups listed by the FDA (Form FDA 2438g (10/91) (Citrus, Pome, Stone,Tropical/Subtropical, Vine Fruits and Small Fruits and Berries) such aswatermelon, white grape, pineapple, lychee, cantaloupe, banana, orange,apple, pear, lemon, passion fruit, red grape, blueberry, tamarind,peach, papaya, acai, plum, guava, tangerine, borojo, cupuacu, goji,kiwi, etc.; this listing is not intended to limit the fruits that aresuitable.

The co-process vegetable, vegetable juice, puree, juice concentrate, drypowder, or extracts may be obtained from vegetables included within thegroups listed by the FDA (Form FDA 2438g (10/91) (Root and Tuber, Bulb,Leaf & Stem, Brassica, Legume, Fruiting and Curcubit Vegetables) such asbean sprouts, green cabbage, celery, onions, sweet onions, asparagus,any leaf vegetables, green beans, peas, cauliflower, broccoli, carrot,pumpkins, bell peppers, potato, sweet potato, tomato, etc.; this listingis not intended to limit the vegetables that are suitable.

The co-process grain powders, grain solutions, paste, or extracts may beobtained from grains included within the groups listed by the FDA (FormFDA 2438g (10/91) (Cereal Grains) such as wheat, barley, rice, oats,corn, sorghum, millets, rye, buckwheat, triticale, fonio, and quinoa,etc.; this listing is not intended to limit the grains that aresuitable.

The co-process oilseeds, nuts, and seeds powder, solution, paste,extracts or derivatives may be obtained from such materials includedwithin the groups listed by the FDA (Form FDA 2438g (10/91) (LegumeVegetables, Oilseeds, Tree Nuts) such as soybean, red kidney beans, limabeans, lentils, chickpeas, black-eye peas, black beans, fava beans,adzuki beans, anasazi beans, peanut, almonds, beechnut, brazils,walnuts, hazels, pistachio, cashews, macadamia, chestnuts, pecan,coconuts, pine nuts, pumpkin seeds, sesame seeds, sunflower seeds,cottonseed, etc.; this listing is not intended to limit the legumes,seeds and nuts that are suitable.

The co-process plant materials may be barley plant extract, grassextracts, tea and tea extracts, seaweeds and extracts etc.; this listingis not intended to limit the plant materials that are suitable.

The co-process animal materials may be skeletal muscle, non-muscleorgans, skins, or shell extracts from mammal animals, poultry, seafood,reptiles, and milk and eggs; this listing is not intended to limit theanimal materials that are suitable.

The co-process materials may be various microbial materials includingyeast or yeast extracts, fungi and fungal extracts including mushroomsand algae and algae extracts etc.; this listing is not intended to limitthe microbial materials that are suitable.

The co-process materials may be the extracts from any of the abovesources and may comprise amino acids, polypeptides, proteins, orcompounds with one or more primary amine groups. Extract solvents may beselected from those common in the art such as de-ionized water;phosphate, or citrate, or acetate, or carbonate etc. buffer solutions,alcoholic solutions or mixtures of the above extractants in differentratios.

Preparation Process for Stable Colorants

In order to produce the colorants of the present disclosure, the matureGenipa americana are co-processed with other fruits such as watermelonby conventional methods well known in the art in order to extract juicefrom fruit. After washing and/or blanching, the fruit is peeled and cutup into pieces, then milled or blended with co-process materials,extracted with or without heating, the pulp, seeds and skin material isseparated by filtration, centrifugation or pressing and the clarifiedjuice is collected. Processing may take place for up to about 8 hours,preferably about 0.1 to 4 hours, and more preferably about 0.1 to 1hours, at a temperature of about 20-45° C. The composite juice obtainedis subjected to subsequent heating to a higher temperature of about50-95° C. for about 1 to 4 hours with suitable mixing (e.g., by shaking,agitation, or aeration) and concentrated about 2 to 10-fold using lowpressure evaporation; the processing has the advantage of providing therequired Juice HACCP treatment for microbial stability. Concentrationalso can be performed on a flash evaporator, by reverse osmosis, orultra-filtration with a suitable membrane, thus providing a concentrate(typically a two- to ten-fold concentration factor from the expelledjuice) suitable for commercial shipping, and the standardization of theresultant color at a stable and desired hue and color strength which maybe adjusted by selecting the ratio of the Genipa americana fruits toco-process materials. The resultant color may subsequently be dried byspray drying, freeze drying or vacuum drying. Alternatively, the processof color manufacture may commence with a previously prepared Huito juiceor concentrate to which the co-process material is added.

This ratio of Genipa americana fruits to co-process materials may bevaried from about 1:0.2 to about 1:30 (by weight) depending on thequantity of amino acids and proteins in the co-process materials, andalso the type or the profile of the amino acids. Specifically, whenGenipa americana fruit is co-processed with fresh watermelon, the ratioof fruit to watermelon is from about 1:1 to about 1:24, and preferablyfrom about 1:2 to about 1:12, and more preferably from about 1:3 toabout 1:6.

Heating in the process relates to enzyme reactions, color development,and color stability. Without wishing to specify a reaction mechanism, itis believed that genipin, or genipin gentiobioside and geniposidic acidthat are hydrolyzed to genipin and genipin derivatives by heating or theaction of beta-glucosidases that exist naturally in the Genipa americanafruits, reacts with proteins and amino acids in the fruits to producethe resultant color (Paik, Y.; Lee, C.; Cho, M.; and Hahn, T. in J.Agric. Food Chem. 2001, 49, 403-432). The overall reaction proceedsslowly at low or ambient temperature. To accelerate the reaction, thecurrent disclosure uses two heating steps. First, low temperatureheating is executed at about 4-about 50° C., and preferably at about20-about 45° C. Most enzymes are active in this temperature range,including proteases which degrade protein to free amino acids,pectinases and cellulases which break down pectin and cellulose. Lowtemperature heating facilitates the release of chemical compounds fromthe cells into the aqueous solution. Second, high temperature heating ispreferably at about 50-about 95° C., more preferably at about 55-about90° C., and most preferably at about 60 to about 85° C., with mixing.During this step, the activity of beta-glucosidase is maximal, and therate of the chemical reaction of genipin with primary amine groups inamino acids, polypeptides or proteins is also high. A significant colorchange can be observed. Heat may also control unwanted reactions. Forexample, the endogenous enzymes polyphenol oxidase and peroxidase whichcause unwanted color and flavor changes in fruits and vegetables afterharvesting and during processing, can be denatured by judicious heating.In the present process, the heating regime contributes to areproducible, stable and desired resultant color.

Process additives may be added in the process. Enzymes, such asbeta-glucosidase or proteases, accelerate color development; otherenzymes, like cellulase, hemi-cellulase, and pectinase, increase totalmass yields of juice.

The pH in the inventive process affects color development. To produce astable blue color, the suitable pH of Genipa americana fruit juice andco-process materials, e.g. watermelon juice, may be from acid to basic,in the range from pH about 3 to about 8, preferably from pH about 3.5 toabout 7, and more preferably from pH about 4 to about 6. To produce astable reddish color, the pH of Genipa americana fruit juice is adjustedto greater than pH about 10, preferably pH greater than about 12. Underalkaline conditions, the iridoid compounds are hydrolyzed and lose themethyl group from the —COOCH₃ group at the C-4 position, resulting in a—COO⁻ group at C-4 position (U.S. Pat. No. 4,247,698 ‘Red coloringcomposite and the method for its production’ Toyama, et al.). Afterholding at alkaline conditions for a sufficient period and at a suitabletemperature, the pH is adjusted back to pH about 3-about 6, and thejuice is co-processed with other materials that contain amino acids,polypeptides, or proteins to generate a reddish, or a reddish-purplecolor.

Other components in the co-process materials, such as antioxidants,multi-ionic metals, reducing sugars, sulfur-containing compounds andpolyphenols, may take part in side reactions during the process.Therefore the color of natural stable colorants varies by change ofco-process materials and processes, with a maximum absorption fromwavelength 400 to 800 nm. More specifically, a brilliant blue generatedby the Genipa americana fruit with watermelon has a maximum absorptionat 585-600 nm wavelength, while a green color generated by Genipaamericana fruit with pineapple juice has absorption maxima at 590-610 nmand 400-420 nm.

The colored solids or water insoluble materials that are obtained duringfiltration or centrifugation steps may also be utilized as valuablecolorants.

Properties of Natural Stable Colorants

Natural colorants that are produced by using Genipa americana andco-processed edible materials have maximum absorptions varying from 360to 800 nm. Specifically, the brilliant bluish colorants that areproduced from Genipa fruit juice and watermelon juice have a maximumabsorption in a spectrophotometer (Perkin Elmer UV/VISspectrophotometer, Lambda 20, USA) in the wavelength range from 585 to600 nm, depending on the concentration of the reactants. The Lab-valuesthat were determined in a Hunter Color Lab calorimeter (Color Quest XE,USA) are L from 20 to 40; a-value from 5 to −2; and b-value from −5 to−25 for concentrated colorants with a color value of 2.0-10.0.

Heat stability of commonly used food colorants is very critical forapplications. The natural colorants generated from this disclosureresist heat very well. After boiling at pH 3 for 30 minutes, Genipa bluecolor obtained with watermelon juice or fresh watermelon shows nosignificant visual changes

Foods and Drink Products Containing Color Materials

Genipa-based natural colorants, especially blue color, have excellentthermal and acidic pH stability, and hence are particularly suitable tocolor foodstuffs, drugs, nutritional supplements, personal care stuffs,cosmetic, and animal feed applications to replace synthetic pigments ordyes.

Genipa-derived natural colorants may be used in foods and drugapplications after appropriately concentrating or after diluting witheither aqueous liquids or suitable approved solvents, including alcohol.Depending on the specific use, Genipa-derived natural colorants, eitheralone or mixed with other dye or pigments, may be used in a broad rangeof foodstuff applications including but not limited to various beveragesand drinks, breakfast cereals, bakery products, pasta/noodles,confectionery, dairy products, processed meat, poultry, and seafoodproducts, different dressings, ice creams, pickles, crackers, and so on.

The following examples are provided for the purpose of furtherillustrating the present disclosure but are in no sense to be taken aslimiting,

EXAMPLE #1

Several frozen and thawed Huito fruits were peeled and cut into smallpieces. One part of fruit, 116.0 g, was mixed with two parts ofde-ionized water, 232.0 g, and blended using a high speed LaboratoryBlender (Waring® Commercial) for 2 periods of 40 seconds each. Then theblended fruit puree was filtered through a No. 4 Whatman filter paper,and the filtered liquid was collected. Beta-glucosidase (EnzymeDevelopment Corp.) was added to the filtrate at a quantity of 0.10 g per60.0 g solution.

To five grams of the solution above in a test tube (25 ml), five gramsof liquid or powder materials from fruits, vegetable, milk, soy, andmeat were added in the test tube, and mixed well. Samples were left tostand at room temperature for 16 hours, and then heated to 80° C. for0.5-2.5 hours until the color was stable. Samples were clarified byfiltration, if necessary, to remove insoluble material.

Color changes of co-processed materials before and after incubation weremeasured in a Hunter Lab calorimeter (Color Quest XE, USA), and resultsare shown in Table 1.

TABLE 1 Before Reaction Heating After Reaction Co-Process Materials L ab hours L a b Visual Color Watermelon concentrate, 26.83 7.13 2.92 2.024.67 0.11 −1.38 Dark Blue 65 Brix Pineapple juice 34.46 5.41 10.16 2.525.96 −0.35 −1.14 Forest green Lychee juice concentrate, 59.94 0.53 8.292.0 24.70 0.04 −1.40 Vibrant blue 29 Brix Passion fruit clarified, 50Brix 25.90 6.15 1.61 2.0 24.65 0.19 −0.95 Dark brown Peach juiceconcentrate, 23.55 0.62 −0.60 2.0 24.54 0.13 −1.04 Black 68 BrixCantaloupe juice 33.66 5.24 6.48 0.5 24.58 0.03 −1.09 Dark blue/puipleBanana puree 2.5 31.39 −0.35 −1.51 Grayish blue Green bean sproutsolution 44.53 0.64 7.79 0.5 24.59 0.06 −1.22 Dark blue/purple Celeryjuice 29.69 9.22 5.30 1.0 24.62 0.04 −1.24 Dark blue Green cabbagepowder 2.0 24.56 0.04 −1.14 Purplish blue Sweet yellow onion solution40.00 −1.46 7.17 1.0 25.33 0.18 −1.38 Grayish purple Milk, 2% 87.80−2.13 7.37 2.0 29.00 −1.12 −6.96 Creamy bright blue Soy milk 79.17 0.138.33 2.0 27.92 −0.97 −4.44 Creamy teal blue Chicken meat slurry 74.960.62 9.22 2.0 24.59 0.09 −1.03 Bright purplish blue

EXAMPLE #2

Peeled and diced Huito fruit, 150.0 g were mixed with fresh dicedwatermelon, 1200.0 g, and pureed for one minute in a blender. Then thepuree/juice was heated in a water bath at 40° C. for one hour. Thepuree/juice was then centrifuged at 2800 rpm for 20 min, and thesupernatant was filtered through fluted filter paper. Next the filtratewas heated in a hot-water bath (80° C.) for 1.5 hours; after which thesolution was blue in color. The blue solution then was concentrated in aRotavapor (Buchi, Switzerland) at 40 mmHg pressure and 50° C. and 100rpm rotation. The final sample that was obtained after 7.5×concentration had an apparent Brix of 75-78, and a maximum absorption of0.13366 at 595.79 nm in a spectrometer (Perkin Elmer UV/VISSpectrometer, Lambda 20, USA) after 100× dilution with de-ionized water.

EXAMPLE #3

The blue colored product was prepared from 2,931 g of peeled Huitofruits with fresh watermelon, 11,305 g, and pureed one minute in ablender. The puree/juice was stirred in a 20 L kettle at about 37-40degree ° C. for one hour after which it was filtered to remove solids bypassage through a number #20 mesh size screen and a 5 μm filter at apump speed of 1.0 liter per minute. The filtrate was returned to thecleaned kettle and heated to about 75-80° C. for 1.5 hours with vigorousstirring. The resultant blue solution was concentrated in a Rotavapor(Buchi, Switzerland) at 40 mmHg pressure and 50° C. and 100 rpmrotation. The final sample had an apparent concentration of 63.25 Brix,and a maximum absorption of 0.57124 at 591.76 nm wavelength in aspectrometer (Perkin Elmer UV/VIS Spectrometer, Lambda 20, USA) after1000x dilution with de-ionized water. The process is diagramed in FIG.1.

EXAMPLE #4

Stability test 1 for low pH beverage applications. An 8 week acceleratedshelf-life stability was performed on three typical beverageformulations: a dairy-based beverage, a vitamin-enhanced water, and alemonade beverage containing approximately 10% juice. The pH of theformulations was between 2.8 and 3.2, and the use rate of the colorpreparation of Example 3 was 0.05%. The accelerated condition was a 90°F. hot box, while a refrigerated 40° F. cold box sample served as thecontrol. Samples were visually evaluated and monitored for color changeusing a Hunter Color Lab calorimeter (Color Quest XE, USA). CalculatedDEcmc values represent the change in color compared to the control basedon L a b data.

After 8 weeks, the dairy-based formula exhibited an approximate colorfade of 20-25% with a change from light blue to a more grayish blue. Thelemonade formula showed an approximate color fade of 30-35% withbrowning of 20-25% and a change from a grayish blue to green. This wasmost likely due to browning of the juices which when combined with theblue color resulted in a shade of green.

After 8 weeks the enhanced water formula demonstrated an approximatecolor fade of 20-25% with browning of 20% and a slight change towardsgreen. Light stability testing (Atlas Suntester XLS+, window glassfilter, E 765 w/m²) consisting of a 6 hr accelerated light exposureusing simulated daylight showed less than 30% fading in all threebeverage formulations.

TABLE 2 8 Week Accelerated Stability 6 Hour Light Stability BeverageDEcmc (RSIN Mode) DEcmc (RSIN Mode) Dairy Base 2.90 0.93 Lemonade 4.811.22 Enhanced water 4.00 2.39

Stability test 2 for low pH beverage applications. An additionalaccelerated test was performed on a beverage containing the blue colormade from a more concentrated color juice batch. The beverage was asimple sugar-acid tasting solution. This beverage only experienced aslight fade of approximately 5% after 8 weeks. Light stability testingshowed very little color change whatsoever.

TABLE 3 8 Week Accelerated Stability 6 Hour Light Stability BeverageDEcmc (RS IN Mode) DEcmc (RSIN Mode) Sugar-acid tasting 1.30 0.43solution

Stability test 3 for low pH beverage applications. A pilot scale-up ofthe blue color using locally sourced materials was set up in a simplesugar-acid tasting solution. After 8 weeks, a fade of approximately 20%was observed. Light stability testing showed a color change ofapproximately 25-30%.

TABLE 4 8 Week Accelerated Stability 6 Hour Light Stability BeverageDEcmc (TTRAN Mode) DEcmc (TTRAN Mode) Sugar-acid tasting 3.37 4.17solution

Stability test 4 for low pH beverage applications. A larger commercialpreparation of the blue color was set up in an enhanced water beverage.This was set up both with and without ascorbic acid (approximately 100%RDI Vitamin C). After 8 weeks, a fade of approximately 30-35% wasobserved in the sample with Vitamin C. This also had a color shifttowards a teal blue-green. The sample without Vitamin C showed fading ofapproximately 20-25% with a color shift towards a more purple blue. Theeffect of Vitamin C on color stability is similar to what would beexpected for other colors from nature such as those which areanthocyanin based. Light stability testing showed a color change ofapproximately 30% in the formulation without Vitamin C and less than 30%in the formulation with. In general, in stability tests, color fade andchanges in hue were within acceptable limits for commercial products andof similar magnitudes to those commonly known for anthocyanin foodcolorants.

TABLE 5 8 Week Accelerated Stability 6 Hour Light Stability BeverageDEcmc (TTRAN Mode) DEcmc (TTRAN Mode) Enhanced Water 7.85 3.13 withVitamin C Enhanced Water 3.53 5.21 without Vitamin C

EXAMPLE #5

Application test 1. The blue color was added to a typical ice creambase. The resultant color in application was a light blue shade.

Application test 2. The blue color was added to a cracker base and thenbaked in an oven. The resultant color in application was a medium bluecolor with a slight greenish/brown tint. The blue color was also addedto a typical cereal base. After going through a commercial extrusionprocess, the resultant color was a grayish blue. These colors wereexamined after 6 months ambient storage and showed little fading.

Application test 3. The blue color was added to a gummy base as well asa pectin base. The resultant color was an attractive deep blue. The userate was lowered in the gummy base to provide a lighter shade of blue.Blending with additional colors from nature provided attractive shadesof green and purple in the gummies.

EXAMPLE #6

Application test. The blue color was added to a cracker base and thenbaked in an oven. The resultant color in application was a medium bluecolor with a slight greenish/brown tint. The blue color was also addedto cereal base. After going through a commercial extrusion process theresultant color was a grayish blue.

EXAMPLE #7

The filtered Huito fruit juice was obtained by the procedure of Example#1, but without the addition of beta-glucosidase. 20.0 g of the juicewas mixed with a 5.0 g of 10% NaOH solution. The mixture was heated to35-40° C., and allowed to stand at room temperature for about 3 hours.The pH of the mixture was adjusted to pH 4.3 by adding 2.08 g of citricacid and 0.04 grams of beta-glucosidase was then added to the solutionand mixed well.

Five grams of the above solution were added to a test tube (25 ml), andanother five grams of liquid test materials from fruits, vegetables, orde-ionized water, were added to the test tube, and mixed well. Sampleswere heated to 80-90° C. for 2 hours until the color was stable. Allsamples showed colors ranging from reddish to brown. Samples werefiltered if necessary.

The absorption maximum wavelength for each of the above colorants weremeasured in a spectrometer (Perkin Elmer UV/VIS Spectrometer, Lambda 20,USA), and the results are shown in Table 6.

TABLE 6 Test Materials added λ max Visual Color Watermelon concentrate,531.82 Reddish brown 65 Brix Cantaloupe juice 540.32 Reddish brown Greenbean sprout solution 528.66 Reddish brown Celery juice 538.03 Reddishbrown Sweet yellow onion solution 534.17 Reddish brown De-ionized water(control) 576.06 Reddish brown

EXAMPLE #8

Gardenia juice, six grams, was added to a test tube (25 ml). Then 0.20 gof beta-glucosidase and 0.10 g of 10% w/w NaOH solution were added tothe test tube, and mixed well. The pH was thereby adjusted from 3.99 to4.60. The Gardenia juice obtained was heated in a 60° C. water bath forone hour, then watermelon concentrate, five grams, was added and mixedwell. The mixture sample was heated to 70° C. and held for 1 hour. Thecolor of the sample was converted from red to green.

EXAMPLE #9

A blue color juice concentrate was prepared by mixing 49.4 g of dicedHuito fruit with 50.0 g of pumpkin juice concentrate (Diana Naturals)and 150 g of de-ionized water. The mixture was blended at a high speedfor 2×60 seconds. Then the puree/juice was heated on a heating plate toa temperature of 42° C. and held for 30 minutes, and then wascentrifuged at 3500 rpm for 15 min, and the supernatant was filteredthrough No. 4 Whatman filter paper. Next the filtrate was heated on aheating plate at a temperature of 60° C. and held for one and a halfhours. A desirable blue color was generated with L value 25.68, a-value0.09, and b value −3.26, measured on a Hunter Colormeter. The bluesolution then was concentrated in a Rotavapor (Buchi, Switzerland) at 40mmHg pressure and 50° C. and 100 rpm rotation. The final sample that wasobtained after 7.3× concentration had an apparent Brix of 67.13.

EXAMPLE #10

A blue/green color juice was prepared with Huito fruit and yellow bellpepper juice concentrate. Fifty grams of yellow bell pepper juiceconcentrate (Diana Natural) were mixed with 250 g of de-ionized water.The solution was heated to boiling and filtered to remove flocculatethrough a No.4 Whatman filter paper. The filtrate was then mixed with64.5 g of diced Huito fruit, and the mixture was blended at a high speed(Waring blender) for 2×60 seconds. A preheating step took place on aheating plate at 42° C. for 30 minutes. After removal of insolublematerial by centrifugation at 3500 rpm for 15 min and filtration throughNo. 4 Whatman filter paper, the filtrate was heated on a heating plateat a temperature setting of 62° C. for one hour. A blue/green color wasgenerated with L value 15.87, a-value −2.85, and b value −6.26, measuredwith a Hunter Colormeter. The blue/green solution then was concentratedin a Rotavapor (Buchi, Switzerland) at 40 mmHg pressure and 50° C. and100 rpm rotation. The final sample that was obtained after 10×concentration had an apparent Brix of 65.2.

CITED DOCUMENTS

-   Endo, T. and Taguchi, H. The constituents of Gardenia jasminoids:    Geniposide and genipin-gentiobioside. Chem. Pharm. Bull. 1973-   U.S. Pat. No. 4,878,921, Kogu et al, issued Nov. 7, 1989-   Sampaio-Santos and Kaplan, J. Braz. Chem. Soc. 12 (2001)-   Andreason and Bremer, Am. J. Botany 87 1731-1748 (2000)-   Form FDA 2438g (10/91)-   Paik, Y; Lee, C.; Cho, M.; and Hahn, T. in J. Agric. Food Chem.    2001, 49, 403-432.-   U.S. Pat. No. 4,247,698, Toyama et al, Jan. 27, 1981

What is claimed is:
 1. Method of preparing stable, natural colors, themethod resulting in a color having an increased -b value based on theCIE LAB scale when the mixture defined in step (a) is compared with theprocessed mixture defined in step (c), comprising a. forming a mixturecomprising: (i) juice from fruit of a plant of the Rubiaceae Family, thefruit comprising juice which contains sufficient genipin or derivativesof genipin, selected from genipin gentiobio side, geniposide,geniposidic acid, and gardenoside, capable of reacting with the juice orliquefied material defined in (a)(ii) to produce a product of thedesired color; (ii) other juice or liquefied material made by thechemical or mechanical liquification of a solid material, from asuitable food-grade source selected from fruits, grains, seeds, beans,nuts, vegetables, plant materials, milk, dairy products, egg, meat,seafood, shellfish, microbial and algal material, and by-products fromsuch sources, the other material comprising juice or liquefied materialmade by chemical or mechanical liquification of a solid material thatcontain components capable of providing the desired color when combinedwith the juice defined in (a)(i); b. processing said mixture usingconventional juice processing methods selected from milling, blending,mixing, pressing, extracting, and combinations of those processingmethods, at a pH of from about 3.5 to about 7; and c. stabilizing themixture against microbial growth, oxidation, organoleptic deteriorationand to provide a stabilized color intensity, by applying a heattreatment.
 2. Method according to claim 1 in which the genipin sourceplant of the Rubiaceae family is Huito, Genipa americana.
 3. Methodaccording to claim 1 in which the genipin source is selected from thewhole fruit, fruit juice, fruit puree, fruit juice concentrate, driedpowder form of fruits or juice, and water insoluble part of fruitsderived, in whole or in part, from plant of the Rubiaceae family isHuito, Genipa americana, its varieties or relatives in the Gardeniagenus, and combinations thereof.
 4. Method according to claim 1 in whichthe genipin source plant of the Rubiaceae family is Gardenia jasminoidesEllis, selected, in whole or in part, from the whole fruit, fruit juice,fruit puree, fruit juice concentrate, dried powder form of fruits orjuice, and water insoluble parts of fruits derived, in whole or in part,from Gardenia jasminoides Ellis, its varieties or relatives in theGardenia genus, and combinations thereof.
 5. Method according to claim1, wherein food grade source (a)(ii) is selected from concentrates,purees and dried forms, and combinations thereof.
 6. Method according toclaim 1, wherein food grade source (a)(ii) is selected from watermelon,white grape, pineapple, tamarind, green cabbage, carrot, orange, apple,pear, red grape, blueberry, bean sprout, cantaloupe, lychee, peach,papaya, lemon, acai, plum, guava, passion fruit, tangerine, borojo,cupuacu, banana, pumpkin, bell peppers, and combinations thereof. 7.Method according to claim 6, wherein food grade source (a)(ii) iswatermelon, and the resulting color is blue.
 8. Method according toclaim 1, wherein the processing includes pH adjustment.
 9. Methodaccording to claim 1, wherein food grade source (a)(ii) containsnitrogenous compounds selected from amino acids, polypeptides, proteins,and compounds with one or more primary amine groups, and combinationsthereof.
 10. Stable natural juice-based colorant made according to themethod of claim
 1. 11. Stable natural juice-based colorant madeaccording to the method of claim
 2. 12. Stable natural juice-basedcolorant made according to the method of claim
 4. 13. Stable naturaljuice-based colorant made according to the method of claim
 7. 14. A foodproduct comprising the colorant of claim
 10. 15. A food productcomprising the colorant of claim
 11. 16. A food product comprising thecolorant of claim
 13. 17. A food product comprising the colorant ofclaim
 12. 18. The method according to claim 1 wherein step (b) takesplace at a pH of from about 4 to about
 6. 19. The method according toclaim 1 wherein the color prepared is blue.
 20. The method according toclaim 19 wherein the material used in (a)(ii) is a juice.
 21. The methodaccording to claim 19 wherein juice from Genipa americana is reactedwith watermelon juice.
 22. A method of preparing stable, natural colors,the method resulting in a color having an increased -b value based onthe CIE LAB scale when the mixture defined in step (a) is compared withthe processed mixture defined in step (c), comprising a. forming amixture comprising: (i) fruit of a plant of the Rubiaceae Family, thefruit comprising juice which contains sufficient genipin or derivativesof genipin, selected from genipin gentiobioside, geniposide, geniposidicacid, and gardenoside, capable of reacting with the juice or liquefiedmaterial defined in (a)(ii) to produce a product of the desired color;(ii) other material from a suitable food-grade source selected fromfruits, grains, seeds, beans, nuts, vegetables, plant materials, milk,dairy products, egg, meat, seafood, shellfish, microbial and algalmaterial, and by-products from such sources, the other materialcomprising juice or liquefied material made by chemical or mechanicalliquification of a solid material that contain components capable ofproviding the desired color when combined with the juice defined in(a)(i), wherein the food-grade source comprises an amino acid; b.processing said mixture using conventional juice processing methodsselected from milling, blending, mixing, pressing, extracting, andcombinations of those processing methods, at a pH of from about 3 toabout 8; and c. stabilizing the mixture against microbial growth,oxidation, organoleptic deterioration and to provide a stabilized colorintensity, by applying a heat treatment.
 23. The method according toclaim 22 in which the plant of the Rubiaceae family is Huito, Genipaamericana.
 24. The method according to claim 22 in which the plant ofthe Rubiaceae family is Huito, Genipa americana, its varieties orrelatives in the Gardenia genus, and combinations thereof.
 25. Themethod according to claim 22 in which the plant of the Rubiaceae familyis Gardenia jasminoides Ellis, its varieties or relatives in theGardenia genus, and combinations thereof.
 26. The method according toclaim 22, wherein food grade source (a)(ii) is selected fromconcentrates, purees and dried forms, and combinations thereof.
 27. Themethod according to claim 22, wherein food grade source (a)(ii) isselected from watermelon, white grape, pineapple, tamarind, greencabbage, carrot, orange, apple, pear, red grape, blueberry, bean sprout,cantaloupe, lychee, peach, papaya, lemon, acai, plum, guava, passionfruit, tangerine, borojo, cupuacu, banana, pumpkin, bell peppers, andcombinations thereof.
 28. The method according to claim 27, wherein foodgrade source (a)(ii) is watermelon, and the resulting color is blue. 29.The method according to claim 22, wherein the processing includes pHadjustment.
 30. A stable natural juice-based colorant made according tothe method of claim
 22. 31. A stable natural juice-based colorant madeaccording to the method of claim
 23. 32. A stable natural juice-basedcolorant made according to the method of claim
 25. 33. A stable naturaljuice-based colorant made according to the method of claim
 28. 34. Afood product comprising the colorant of claim
 30. 35. A food productcomprising the colorant of claim
 31. 36. A food product comprising thecolorant of claim
 32. 37. A food product comprising the colorant ofclaim
 33. 38. The method according to claim 22 wherein step (b) takesplace at a pH of from about 4 to about
 6. 39. The method according toclaim 22 wherein the color prepared is blue.
 40. The method according toclaim 39 wherein the material used in (a)(ii) is a juice.
 41. The methodaccording to claim 39 wherein juice from Genipa americana is reactedwith watermelon juice.
 42. The method according to claim 22 wherein theamino acid is a natural amino acid.
 43. The method according to claim 22wherein step (b) takes place at a pH of from about 3.5 to about
 7. 44.The method according to claim 22 wherein the color prepared is teal blueor purple blue.
 45. A method of preparing stable, natural colors, themethod resulting in a color having an increased -b value based on theCIE LAB scale when the mixture defined in step (a) is compared with theprocessed mixture defined in step (c), comprising a. forming a mixturecomprising: (i) Genipa americana fruit juice which contains sufficientgenipin or derivatives of genipin, selected from genipin gentiobioside,geniposide, geniposidic acid, and gardenoside, capable of reacting withthe juice or liquefied material defined in (a)(ii) to produce a productof the desired color; (ii) other juice or material from a suitablefood-grade source that contains at least one component capable ofproviding the desired color when combined with the Genipa americanafruit juice defined in (a)(i), wherein the food-grade source comprisesan amino acid; b. processing said mixture using conventional juiceprocessing methods selected from milling, blending, mixing, pressing,extracting, and combinations of those processing methods, at a pH offrom about 3 to about 8; c. stabilizing the mixture against microbialgrowth, oxidation, organoleptic deterioration and to provide astabilized color intensity, by applying a heat treatment; and d. dryingthe mixture, wherein the drying is selected from the group consisting ofspray drying, freeze drying, and vacuum drying, and combinationsthereof.
 46. The method according to claim 45 wherein the heat treatmentcomprises heating the mixture for a predetermined period of time. 47.The method according to claim 46 wherein the predetermined period oftime is about 0.1 to 8 hours.
 48. The method according to claim 46wherein the predetermined period of time is about 0.1 to 4 hours. 49.The method according to claim 46 wherein the predetermined period oftime is about 0.1 to 1 hours.
 50. The method according to claim 45wherein the heat treatment comprises heating the mixture at about 50-95degrees Centigrade for a predetermined period of time of about 1 to 4hours.
 51. The method according to claim 45, wherein food grade source(a)(ii) is selected from concentrates, purees, extracts, pastes anddried forms, and combinations thereof.
 52. The method according to claim45, wherein the color is blue.
 53. A stable natural juice-based colorantmade according to the method of claim
 45. 54. A stable naturaljuice-based colorant made according to the method of claim
 50. 55. Afood product comprising the colorant of claim
 53. 56. A food productcomprising the colorant of claim
 54. 57. The method according to claim45 wherein step (b) takes place at a pH of from about 3.5 to about 7.58. The method according to claim 45 wherein step (b) takes place at apH of from about 4 to about
 6. 59. The method according to claim 45wherein the Genipa americana fruit juice is obtained by peeling Genipaamericana fruit from its skin to obtain a skinless Genipa americanafruit, and obtaining the Genipa americana fruit juice from the skinlessGenipa americana fruit.
 60. The method according to claim 59 wherein theobtaining the Genipa americana fruit juice from the skinless Genipaamericana fruit comprises milling or pureeing the skinless Genipaamericana fruit.
 61. The method according to claim 60 wherein theobtaining the Genipa americana fruit juice from the skinless Genipaamericana fruit further comprises processing selected from the groupconsisting of filtering, centrifuging, or pressing the milled or pureedskinless Genipa americana fruit.
 62. The method according to claim 61wherein the processing comprises filtering or pressing the milled orpureed skinless Genipa americana fruit against a mesh.
 63. The methodaccording to claim 1 further comprising drying the mixture wherein thedrying is selected from the group consisting of spray drying, freezedrying, and vacuum drying, and combinations thereof.
 64. The methodaccording to claim 22 further comprising drying the mixture wherein thedrying is selected from the group consisting of spray drying, freezedrying, and vacuum drying, and combinations thereof.